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LDL& HDL

This is hetrogeneous group of compounds which are important constituents
of all cells the nerve cell are highly enriched with lipids
The lipids are insoluble water and therefore present in serum either in
combination with certain proteins (Lipoproteins) or in an emulsified form as
chylomicrons.
Plasma lipoprotein are classified into 4 major groups according to density
1. hylomicrons composed of trigycerides! Transport trigycerides from
the gut
". #L$L (very low density lipoproteins) composed of trigycerides! 1%&
Transport trigycerides from liver to peripheral tissues.
'. L$L (low density lipoproteins) ()& composed of cholesterol esters.
Transport cholesterol to tissues
4. *$L (*igh density lipoproteins) "%& composed of cholesterol esters.
Transport cholesterol from tissues to liver.
CLINICAL PRESENTATION
+stimation of lipoprotein *$L and L$L is playing greater role in
diagnosis, management and prognosis of patients with coronary
artery disease.
-ormal value HDL 35- 75 mg/dl
-ormal value LDL 130-160 mg/dl
Patients suffering from coronary artery disease have low level of *$L
below '%mg.dl and high level of L$L above 1()mg.dl

HDL
OBJECT: determination of high density lipoprotein cholesterol
Principles:
Chylomicrons, VLDL and LDL are precipitated by adding
phosphotngstic acid and magnesim ions to the sample!
Centrifgation lea"es only the HDL in the spernatant!
Reagents:
Concentrations of the reagents
#hospotngstic acid $! mmol%L
&agnesim chloride '!( mmol%L
Procedure: sample / standard 200 ul
Regent 500 ul
Mix and incubate for 5 minute t!en centrifuge for 20 minutes at
2500g" #it!in 2 !ours after centrifugation transfer 0"ml of clear supernatant to
t!e reaction solution for determination of c!olesterol"
$a%elengt! 500nm"

)tandard )ample
)pernatant * $++ ,L
)tandard $+++ , L *
Cholesterol reagent $+++ , L $+++ , L
&i- and incbate for $+ min at room temperatre or . min and at /0
o
C! Then
measre the absorbance of the sample or the standard against the reagent blan1 "ale
2ithin 3. min!
&alculation:
4ith standard
HDL 5 cholesterol 6mg % dL7 8 9 : sample ; conc! )tandard 6mg % dL7
9 : standard
LDL
OBJECT: determination of lo2 density lipoprotein cholesterol
Principle:
Lo2 density lipoproteins <LDL= are precipitated by addition of heparin!
High density lipoproteins <HDL= and "ery lo2 density lipoproteins <VLDL= remain in
the spernatant after centrifgation and are measred en>ymatically by the CHOD*
#:# method! The concentration of LDL cholesterol is calclated as the difference of
total cholesterol and cholesterol in the spernatant!
Reagents:
Concentration of the reagents
Heparin $++ +++ ?%L
)odim citrate (3 mmol%L
Procedure: sample / standard 00 ul
Regent 000 ul
Mix and incubate for 5 minute t!en centrifuge for 20 minutes at
2500g" #it!in !ours after centrifugation transfer 00 ul of clear supernatant to
t!e reaction solution for determination of c!olesterol"
$a%elengt! 500nm"

)tandard )ample
'upernatant * $++ ,L
'tandard $++ , L *
&!olesterol reagent $+++ , L $+++ , L
&i- and incbate for $+ min at room temperatre or . min and at /0
o
C!, read absorbance of
the sample for the standard 2ithin 3. min! against reagent blan1
&alculation:
Cholesterol in spernatant
Cholesterol spernatant 6mg % dL7 8 9 : sample ; conc! )tandard 6mg % dL7
9E standard